Purpose :
Thrombospondins-1 (TSP1) is a matricellular protein which has been shown to regulate cytoskeleton, cell adhesion, and extracellular matrix remodeling. TSP-1 is found in the trabecular meshwork (TM) and is increased in one third of patients with primary open-angle glaucoma (POAG).[1] As TSP1-null mice demonstrated a lower IOP resulting from increased outflow [2], we hypothesized that the suppression of TSP1would alter IOP by affecting extracellular matrix (ECM) synthesis and/or turnover in the trabecular meshwork (TM).

Methods :
A lentivirus carrying the short hairpin RNA targeting human TSP1 (lenti-shTSP1) was constructed using the pLKO.1 vector system. Lenti-shTSP1 was then used to suppress TSP1 in human TM endothelial cells at the multiplicity of infections (MOI) 5. ECM proteins from cell lysates and conditioned media were analyzed by immunoblotting. Human anterior chamber perfusion culture (ex vivo) was treated with lenti-shTSP1 and IOP was recorded every 6 hours.